The present invention relates generally to the separation and/or purification of an enzymatic polyflavonoid with or without a template such as PEO (polyethylene oxide), SPS (sulfonated polystyrene) DBSA (sodium dodecyl benzene sulfonic acid), one or more of the recovered highly pure fractions and the use of one or more fractions. One or more of the recovered fractions can be used as a food antioxidant and/or a medicinal agent such as a drug.
Phenolic polymers, in particular phenol-formaldehyde resins such as novolaks and resols find wide application as wood composite laminates, foundry resins, abrasives, friction and molding materials, coatings and adhesives, fiber bonders and flame retardants. There has been a major concern however over the toxic effects of formaldehyde which are used in current art synthetic processes. In recent years there has been an effort towards the development of an alternative synthetic route which circumvents the toxic effects of formaldehyde. These synthetic routes are also not suitable for forming polymers that must be biocompatible for uses such as food additives.
Enzymatic polymerization of phenol in aqueous solutions has been initially investigated as a possible benign and environmentally friendly solution to the synthesis of polyphenols. These initial attempts however were unsuccessful since they could form only very low molecular weight (oligomers) exhibiting properties insufficient to meet the requirements for engineering applications. Alternative enzymatic reactions have since been investigated to improve molecular weight and the mechanical, thermal, optical and electronic properties of these polymers including micelles, air-water interface monolayers and mixture of various organic solvents. Although the resulting molecular weights and properties of these polyphenols were improved, their commercial application remains limited due to extensive branching and poor solubility and processability of the resulting polymers.
Recently a patent publication by Bruno et al. addresses these problematic issues and results in a new class of polyelectrolyte/polycatechin complexes which are prepared in aqueous medium using an environmentally safe, inexpensive, and facile “one-step” reaction. In addition, these polycatechins are water soluble with improved properties including molecular weight, structural, thermal, antioxidant properties and processability than current synthetic routes. Moreover, the polycatechin were found to exhibit antitumorigenic activity in in vitro studies. Flavonoids are also known to be strong antioxidants and are presently tested as food stabilizers.
However the industrial application of these polymers is hindered by the difficulties in separating the different polymers that are formed in the enzymatic polymerization. The control of structural complexity through controlled polymerization is a technical hurdle. There are 7 possible reaction sites for each flavonoid monomer resulting in 28 possible combinations for the formation of a simple dimer. The possible tetramer structures, formed by two dimers, are 107. Moreover, the procedure for the separation of the different pure fractions is challenging. The structures created from the reaction can be very similar and consequently difficult to separate.